Polysaccharides isolated from traditional Chinese medicines (TCMs) exhibit multiple pharmacological activities. However, quality control of polysaccharides is a challenge because of their complicate structure and macro-molecular mass. In this study, saccharide mapping based on specific enzymatic digestion of polysaccharides and chromatographic analysis was proposed to discriminate the polysaccharides from different TCMs. Endo-carbohydrase such as glucanase, arabinanase, xylanase, galactanase, cellulase, amylase and pectinase were used for enzymatic digestion of polysaccharides from 9 TCMs namely Panax ginseng, P. notoginseng, P. quinquefolium, Cordyceps sinensis, C. militaris, Ganoderma lucidum, G. sinense, Astragalus membranaceus and Angelica sinensis. By using high performance size exclusion chromatography (HPSEC) as well as derivatization with 1-Phenyl-3-methyl-5-pyrazolone (PMP) and HPLC analysis, the enzymatic hydrolysis properties of polysaccharides and their saccharide mapping were determined. The polysaccharides from 9 TCMs were firstly successfully distinguished based on their characteristic saccharide maps, which is helpful to improve the quality control of polysaccharides. © 2009 Elsevier B.V. All rights reserved.